Apparatus for execution of treatment operations on microscope slides with tissue specimens

ABSTRACT

An apparatus for automatic execution of different treatment operations in connection with staining of tissue specimens on microscope slides, wherein the apparatus ( 1 ) comprises an assembly of vessels ( 4 ) for receiving different liquids for staining of the tissue specimens, a loading station ( 2 ) for microscope slides ( 28 ), a conveyor ( 5 ) for transfer of carriers with microscope slides from vessel to vessel in accordance with a treatment program, an unloading station ( 8 ) for treated microscope slides, and a control unit ( 18 ) for controlling the treatment operations in accordance with a data program. The apparatus comprises different levels (I, II) having units for execution of the relevant treatment operations. Thus, a first level (I) comprises the loading station ( 2 ) and the assembly of said vessels ( 4 ) with the appurtenant conveyor ( 5 ) and a second level (II) comprises a station ( 6 ) for application of cover glasses on the stained microscope slides ( 28 ), a succeeding station ( 7 ) for drying of the cover-slipped microscope slides, and the unloading station ( 8 ), a means ( 9 ) being provided for gripping and transfer of carriers ( 10 ) with stained microscope slides from the first level (I) to the cover-slipping station ( 6 ) on the second level (II).

This application is a continuation of U.S. patent application Ser. No.11/793,562, now U.S. Pat. No. 7,998,408, filed Jun. 3, 2008 entitled “AnApparatus for Execution of Treatment Operations on Microscope Slideswith Tissue Specimens” which is a 371 application of PCT/NO2005/000474filed Dec. 22, 2005 and which claims priority to Norwegian PatentApplication 2004 5622 filed Dec. 23, 2004, the disclosures of all areincorporated herein by reference.

The invention relates to an apparatus for automatic execution ofdifferent treatment operations in connection with staining of tissuespecimens on microscope slides, wherein the apparatus comprises anassembly of vessels for receiving different liquids for staining of thetissue specimens, a loading station for microscope slides, a lo conveyorfor transfer of carriers with microscope slides from vessel to vessel inaccordance with a treatment program, an unloading station for treatedmicroscope slides, and a control unit for controlling the treatmentoperations in accordance with a data program.

There are previously known a number of different apparatuses of theabove-mentioned type. As examples of prior art in this field referencecan e.g. be made to U.S. Pat. No. 5,573,727 and U.S. Pat. No. 5,601,650.Both of these publications relate to apparatuses that are restricted tooperations in connection with staining of tissue specimens on microscopeslides. Thus, in the apparatus according to U.S. Pat. No. 5,601,650, themicroscope slides are transported in microscope slide holders by meansof a conveyor to treatment stations wherein they are subjected totreatment steps corresponding to a selectable staining process. Theconveyor is constructed such that each microscope slide holder isseparated therefrom after introduction into a treatment station, so thatother microscope slide holders, in the course of the time a treatmentoperation is carried out at said treatment station, can be transportedto free treatment stations in accordance with the selected stainingprocess.

A main object of the invention is to provide a multi-function apparatuswhich is able to carry out many different operations in connection withtreatment of tissue specimens on microscope slides.

An additional object of the invention is to provide such an apparatuswhich has a relatively compact construction.

The above-mentioned objects are achieved by means of an apparatus of theintroductorily stated type which, according to the invention, ischaracterised in that the apparatus comprises different levels havingunits for execution of the relevant treatment operations, a first levelcomprising the loading station and the assembly of said vessels with theappurtenant conveyor, and a second level comprising a station forplacing of cover glasses on the stained microscope slides, a succeedingstation for drying of the cover-slipped microscope slides, and theunloading station, a means being provided for gripping and transfer ofcarriers with stained microscope slides from the first level to thecover slipping station on the second level.

With the apparatus according to the invention there can for example becarried out initial fixation of paraffin-embedded tissue specimens onmicroscope slides, staining of tissue specimens on microscope slides,mounting of cover glasses on stained tissue specimen microscope slides,and drying of stained tissue specimen microscope slides afterapplication of glue and mounting of cover glasses, in accordance with aselected running process for the apparatus.

In an advantageous embodiment of the apparatus the carrier comprises anelongated hanger having a means for releasable clamping of themicroscope slides in a hanging position with the surfaces located in acommon plane. Such a carrier can be used during execution of all therelevant treatment operations, without manual handling or manipulationof the microscope slides.

In an additional advantageous embodiment of the apparatus according tothe invention, wherein the apparatus comprises a plurality of tanks forthe relevant liquids, wherein the tanks are connected with appurtenantvessels via line connections on which there are arranged pumps for thesupply of liquid to the vessels, the control unit is arranged forselective control of the relevant pumps, so that a vessel to be used ina staining process, is filled with liquid at the start of the stainingprocess, and such that the liquid-filled vessels are emptiedautomatically when they are no longer to be used in the stainingprocess. Such an embodiment implies a particularly efficient utilizationof the relevant treatment liquids. In addition there is achieved asubstantial increase of the time between maintenance of the treatmentliquids, a drastic reduction of the risk that operator personnel issubjected to dangerous fumes from the liquids, and avoidance ofdegradation of the liquids in that they are unnecessarily subjected tosurrounding air.

Additional objects and advantages of the invention will appear from thefollowing further description of exemplary embodiments with reference tothe drawings, wherein

FIG. 1 shows a principle drawing of an apparatus according to theinvention;

FIG. 2 shows a perspective view of a carrier or holding device formicroscope slides, for use in the apparatus according to the invention;

FIGS. 3A and 3B show schematically an embodiment of an arrangement forsupply of liquid from the tanks to the vessels in the apparatus, and forwashing of the vessels with water;

FIG. 4 shows a perspective view seen from above of a vessel arranged forreleasable mounting at a treatment station in the apparatus of FIG. 1;

FIG. 5 shows a perspective view of the vessel in FIG. 4, wherein holdingdevices with microscope slides are placed in the vessel;

FIG. 6 shows a perspective view of a conveyor for use in the apparatusof FIG. 1;

FIG. 7 shows the conveyor in FIG. 6 wherein a holding device withmicroscope slides are suspended from the conveyor; and

FIG. 8 is a perspective view showing the row of staining/reagentstations in the apparatus and the conveyor thereof, and also partly theadditional stations arranged above the reagent stations for treatment ofmicroscope slides with tissue specimens.

As appears from FIG. 1, the apparatus 1 according to the inventioncomprises three different levels, I, II and III, having units forexecution of the relevant treatment operations.

The first level I includes a loading and preheating station 2 formicroscope slides, a number of reagent stations 3 consisting of vessels4 (see FIGS. 4 and 5) for staining of tissue specimens on microscopeslides, and a conveyor 5 for transfer of carriers/holding devices withmicroscope slides from vessel to vessel in accordance with a selectedtreatment program.

The second level II includes a station 6 for application of coverglasses on the stained microscope slides, a succeeding drying station 7and a storing/unloading station 8 for the ready-treated microscopeslides. In the illustrated embodiment the second level II is locatedabove the first level I, and a hoist or lifting means 9 is provided fortransfer of carriers 10 with stained microscope slides from the firstlevel to the cover glass applying or cover-slipping station 6 on thesecond level. A horizontal transport means 11 is provided for transferof carriers 10 with microscope slides through the cover-slipping station6 and the drying station 7 to the unloading station 8.

The third level III includes an assembly 12 of a number of storage tanks13 for the relevant reagent liquids, wherein the tanks are connected toappurtenant vessels 4 at the reagent stations 3 via connecting lines(not shown in FIG. 1) on which there are arranged pumps 14 for thesupply of liquid to the vessels, and moreover filters 15 for filteringof the liquids. The arrangement for liquid supply and for washing of thevessels will be further described in connection with FIGS. 3A and 3B.

The apparatus in FIG. 1 is also shown to include a filter/blower unit 16for ventilation of the apparatus. This unit is in communication with avent inlet 17 at the top of the apparatus, and thereby provides fortransport and blow-out of venting air through the apparatus. In thismanner the whole apparatus is ventilated, so that i.a. dangerous fumesfrom the reagent stations are removed in a justifiable manner.

The apparatus also includes an electronic control unit which issymbolically shown as a block 18, and which takes care of controllingthe treatment operations of the apparatus in accordance with a dataprogram. Further, the apparatus comprises a control panel (not shown)for operation and working of the apparatus. This panel includes a numberof touch or contact keys (control keys and working keys), and also adisplay giving information about the status etc. of the apparatus duringoperation.

As appears from FIG. 1, the vessels 4 at the reagent stations 3 arearranged in a single rectilinear row from the loading station 2 to adedicated area 19 for transfer of carriers 10 with stained microscopeslides from the first to the second level. The conveyor 5 which is to befurther described in connection with FIG. 6, therefore needs to carryout only an xz-movement, i.e. movements only in a horizontal x-directionand a vertical z-direction.

The means 9 for gripping and transfer of carriers is only schematicallyshown in FIG. 1, and consists of a hoist for lifting of the carriersfrom the dedicated transfer area 19 to the second level II. As analternative to such a lifting means, the transfer means may be arrangedat the second level and arranged to fetch carriers 10 from the area 19to the second level.

The station 6 for application of cover glasses on the stained microscopeslides is also only schematically shown in FIG. 1. This station includesa number of elements, more specifically a cover glass magazine with adispenser function, a glue pump, a glue dispenser and a unit formounting of cover glasses on the supplied microscope slides with stainedtissue specimens. Further, the station contains a detection unit fordetection of microscope slides arriving at the station in the differentsupplied carriers. The cover glass magazine advantageously may beconstructed for delivering cover glasses laterally from the bottom ofthe magazine. For a further description of such a station reference maye.g. be made to the international patent application No. PCT/NO99/00396(WO 00/37986).

As appears from FIG. 1, the cover glass applying or cover-slippingstation 6, the drying station 7 and the unloading station 8 are arrangedin succession in a direction which is opposite to the direction from theloading station 2 to the means 9 for transfer of carriers from the firstto the second level. In this manner there is obtained a compactapparatus with a little lateral extension.

As regards the loading and preheating station 2, this comprises a numberof heated stations for fixation of paraffin on tissue specimenmicroscope slides, and in addition a number of liquid stations or emptystations for initial placing of carriers with microscope slides, werethe different stations are used in accordance with the relevant orprogrammed treatment process that is to be used. The station 2 alsoincludes a rush or emergency outlet station which is used for deliveryof a carrier with microscope slides with tissue specimens to be analyzedimmediately after a finished staining operation, without being providedwith cover glasses over the tissue specimens. In such a case the controlunit 18 will be programmed such that the conveyor with the carrier inquestion is directed to the rush output station.

An embodiment of the carrier/holding device 10 which is used in theapparatus according to the invention, is shown in FIG. 2.

As shown, the carrier 10 comprises an elongated hanger which in theillustrated embodiment consists of a carrier plate 25 on which there aremounted a spring-loaded clamp plate 26 which, in cooperation with thecarrier plate, forms a clamp jaw 27 for releasable clamping of a numberof microscope slides 28 in a hanging position with the surfaces of themicroscope slides located in a common plane. In use of the hanger 10with the clamped microscope slides, this is placed in the relevantvessel 4 with its longitudinal direction oriented transversely to thehorizontal transport direction of the conveyor 5 from vessel to vessel.As appears from FIG. 1, the means 9 for gripping and transfer ofcarriers 10 is arranged to place the carrier/hanger with the microscopeslides 28 in a horizontal lying position on the cover-slipping station6. The above-mentioned detection unit on the station 6 can then in asafe manner detect the positions of the microscope slides in thecarriers supplied to the station.

For a further description of the carrier/holding device 10 and itscooperation with a loading rack for loading of microscope slides,reference is made to the simultaneously lo filed patent applicationhaving the title “A holding device for microscope slides with tissuespecimens”.

An embodiment of an arrangement for liquid supply from the tanks 13 tothe vessels 4 and for washing of the vessels with water, is shown inFIGS. 3A and 3B.

In the embodiment shown in FIG. 3A a pump 14 and a filter 15 areconnected on a supply line 30. The pump preferably is of the peristaltictype, and it is preferably reversible, so that used reagent liquid canbe returned to the vessel via the supply line 30. Further, a separatereturn line 31 is arranged between the vessel 4 and the tank 13, and anexhaust or outlet line 32 is arranged between the vessel and a wastetank 33.

The filters 15, which are arranged to secure the quality of the suppliedliquids, are shown to be arranged at the suction side of the pumps 14.However, they can with advantage be located on the pumping side of thepumps, to achieve the best possible pumping efficiency.

As further appears from FIG. 3A, each of the reagent vessels 4 in theapparatus in FIG. 1 is placed in an appurtenant recess 34 in a carrierplate 35 arranged in the apparatus. The vessel 4 in the figure is shownin longitudinal section. The vessel is provided with an inlet opening 36in the bottom of the vessel at one end of the vessel, and with an outletopening 37 in the bottom of the vessel at an opposite side of anoverflow wall 38 in relation to the inlet opening 36, the arrangementbeing such that a through-flow of liquid is produced through the vesselwith continuous supply of liquid to the vessel by means of the relevantpump. The through-flow is obtained in that a perforated spreading plate39 is placed above the bottom of the vessel, so that liquid flowing intothe vessel is distributed over the bottom of the vessel below thespreading plate 39, before the liquid flows through the holes in thespreading plate and thereby causes a through-flow of liquid through thewhole vessel before it flows over the overflow wall 38 and out of thevessel.

The vessels 4 are releasably mounted in the recesses 39 in the carrierplate 35, and the inlet and outlet openings 36 respectively 37 of thevessels are connected in a sealing manner to the connecting lines 30 and31 by means of O-ring seals 40 which are clamped between the undersideof the vessels 4 and the bottom of the recesses 34 when the vessels areput in place in the recesses.

A appears from FIGS. 3A and 3B, a water inlet 41 and a water outlet 42are arranged in the bottom of the recess 34 at the opposite side of therecess in relation to the inlet for the connecting line 30 and theoutlet for the return line 31. The water inlet 41 and the inlet from theconnecting line 30 are arranged symmetrically about a midpoint of thevessel, and the same is the case with the water outlet 42 in relation tothe return line 31. By mounting the vessels 4 in the apparatus in areversed position, i.e. turned 180.degree. relative to the normaloperating position in FIG. 3A, it is thereby achieved that the vesselsare connected automatically to a system for flushing and cleaning of thevessels with water. This position of the vessel 4 is shown in FIG. 3B.Thus, by means of this arrangement the vessels can be flushed andcleaned without being removed from the apparatus.

If the O-ring seals 40 in the recess 34 should not be quite tight,possible leaked-out liquid will be directed from the recess to the wastetank 33 by means of the outlet line 32.

In the embodiment shown in FIG. 3A, the vessel 4 is supplied withreagent liquid from a single appurtenant tank 13. However, a vessel 4may be supplied with liquid from several tanks in that the connectinglines from these tanks are coupled to a multi-way valve connecting thetank containing the relevant liquid. For a further description of suchalternative embodiments reference is made to the simultaneously filedinternational patent application having the title “Method and system foruse of treatment liquids in an apparatus for staining of tissuespecimens on microscope slides”.

A vessel 4 which is arranged for releasable mounting at a stainingstation in the apparatus of FIG. 1, is shown in FIG. 4. The vessel hasan elongated shape and is divided into two parallel spaces 45 by meansof a longitudinally extending partition 46. As mentioned above inconnection with FIG. 3A, each space 45 is provided with an inlet opening36 in the bottom of the vessel at one end of the vessel, and with anoutlet opening 37 in the bottom the vessel at an opposite side of anoverflow wall 38 in relation to the inlet opening 36.

Each of the end walls of the vessel 4 is provided with four notches 47for receiving suspension portions at the ends of carriers/holdingdevices for microscope slides. As mentioned above, these holding devicesare further described in the simultaneously filed international patentapplication having the title “A holding device for microscope slideswith tissue specimens”.

FIG. 5 shows a perspective view of a vessel 4 in which there are placedfour holding devices 48, the suspension portions 49 of the holdingdevices being placed in respective ones of said notches 47. As furtherappears, the vessel at its ends is provided with fastening members 50and 51 for releasable mounting of the vessel at the relevant reagentstation in the apparatus 1. The fastening member 50 is here a fixedmember, whereas the fastening member 51 is resilient, to allow aresilient fixing thereof.

FIG. 6 shows an embodiment of a conveyor 5 for use in the apparatus ofFIG. 1. The conveyor 5 comprises an inverted U-shaped or portal-likemain body 55 consisting of a transverse member 56 extending above andparallel with the reagent stations 3 of the apparatus, and a pair ofside members 57 respectively 58 located at a little distance outside ofthe reagent stations at each end thereof, as shown in FIG. 8. At theinside of each of the side members 57, 58 there is mounted a liftingmeans 59 respectively 60 which is adapted to lift one or two carriers 10with microscope slides. Each lifting means is slidably mounted on a pairof vertical guide columns 61, 62, and can be lifted or lowered on theguide columns by means of a toothed belt 63 running over a lower and anupper toothed belt wheel 64 respectively 65 mounted in the side member57. The lower wheel 64 is driven by a step motor 66. The drive movementis transferred from the upper toothed belt wheel 65 via a transfer shaft67 to a corresponding toothed belt drive (not visible in FIG. 6) mountedon the other side member 58.

The conveyor further comprises a pair of fixing blocks 68 and 69 forhorizontal toothed belt drive of the conveyor, i.e. for theprogram-controlled movement of the conveyor in the x-directiontransversely to the reagent stations 3. This toothed belt drive isschematically shown in FIG. 8 where there are shown a pair of toothedbelt wheels 70, 71 with appurtenant toothed belts 72.

In its horizontal movement the conveyor is guided by a pair of runningwheels 73, 74 mounted on the fixing block 68 on the side member 57 andrunning along a guide rod 75 (see FIG. 8), and a running wheel 76mounted on the fixing block 69 on the side member 58 and running on anangular rail 77.

FIG. 7 shows the conveyor 5 in FIG. 6, wherein a carrier/holding device10 with microscope slides 28 are suspended from the lifting means 59 and60 of the carrier.

The schematic perspective view in FIG. 8 also shows a part of thestations arranged at the second level II above the reagent stations 3and the conveyor 5 at the first level I, more specifically the dryingstation 7 and the storage/unloading station 8. The horizontal transportmeans 11 shown in FIG. 1, is left out in FIG. 8. The figure shows anentry portion 78 of a sliding plate where a carrier 10 with microscopeslides 28 is supplied to a horizontally lying position at the station 6,ready for application of cover glasses on the microscope slides. Thesliding plate extends through the station 6 (not further shown in FIG.8) and the drying station 7 and is terminated at the storage andunloading station 8.

As further appears from the figure, the transport means advances thecarriers 10 with cover-slipped microscope slides through the dryingstation 7 to the station 8 where the carriers are stored for unloadingin a stack wherein the carriers rest partly on each other in a slantingposition (at an angle of about 10.degree. with the horizontal).

A complete operational sequence in operation of the apparatus accordingto the invention will be described below.

At the start of an operational sequence one or more carriers/holdingdevices are loaded into a dedicated area of the loading station,dependent on the types of tissue specimens being on the microscopeslides in the carrier. One area of the loading station may be dedicatedfor fixation of paraffin-embeded tissue specimens on microscope slides,and another area may be liquid vessels for other tissue specimens. Theoperator thereafter will select a program for an inserted carrier.Alternatively the carrier may be able to be started automatically inaccordance with a selected standard program.

After insertion in the loading area the carrier will remain in this areain accordance with a programmed time or according to the finishingintervals of the total lo process. The carrier thereafter is movedautomatically from the loading station to the second step in thetreatment operation by means of the conveyor. The vessel for this stepwill be supplied with liquid from a tank through the pumping systemconnected to the vessel, so that the vessel will be liquid-filled at thetime when tissue specimens on the microscope slides in the carrier areplaced in the vessel. In order to get a good and i efficient treatmentoperation, there will be supplied a flow of liquid which can be adjustedby means of speed adjustment of the motor of the pump. When the timecriterion in this step is satisfied in accordance with a given protocol,the vessel may be emptied and filled with a new liquid in accordancewith the protocol by filling of a new liquid from another connected tankvia a valve connected to the same vessel, or the conveyor may move thecarrier to a new vessel dependent on how the apparatus is set up, inorder to give a shortest possible throughput speed. These transfers ofcarriers in accordance with different protocols and control of thesupply into and out of vessels with one or more reagents, take placecontinuously so that each carrier will have been placed at the correcttime in each liquid in accordance with its protocol. During running theapparatus will register through-flown liquid quantity in connection withthe effective life of the liquid, to thereby be able to give a warningwhen the different liquids are to be replaced. This takes place at thesame time as more carriers can be loaded into the loading station.

When a carrier has been delivered to the dedicated transfer area, thetransfer means of the apparatus will transfer the carrier to thecover-slipping station for application of glue and cover glasses on themicroscope slides detected in the carrier. This takes place at the sametime as carriers can be loaded into the loading station, and at the sametime as other carriers are treated in liquids in accordance with theirprotocol.

By means of the transport means at the second level of the apparatus theready-stained and mounted tissue specimen microscope slides located inthe carrier will be transported through the drying station, so thatsolvent being on the microscope slides/-cover glasses will be driedaway, whereafter the carrier in question will be transported to thedelivery station.

What is claimed is:
 1. An apparatus for automatic execution of different treatment operations in connection with staining of tissues specimens on microscope slides, the apparatus comprising: a plurality of tissue processing stations, at least one of the plurality of tissue processing stations comprising an assembly of tissue processing vessels, each tissue processing vessel configured to receive different reagent for staining of the tissue specimens in accordance with a selected treatment program; a loading station to load the microscope slides carrying tissue specimens onto the apparatus; a conveyor to transport the microscope slides between the plurality of tissue processing station; an unloading station for unloading the cover-slipped microscope slides from the apparatus; and a control unit configured to control the execution of the different treatment operations; wherein the apparatus comprises separate areas and associated devices for execution of the different treatment operations; a first area comprising the loading station and the plurality of tissue processing station; a second area comprising a cover slipping station configured to place cover slips on the microscope slides, a succeeding station for drying the cover-slipped microscope slides, and the unloading station; a hoisting device configured to grip and transport the microscope slides from the first area to the cover slipping station in the second area; and a third area comprising a plurality of reagent storage tanks, wherein each tank is connected via a reagent supply line to at least one of the plurality of tissue processing stations.
 2. The apparatus of claim 1, wherein the plurality of tissue processing stations are arranged in a single rectilinear row from the loading station to a dedicated area for transport of the microscope slides from the first area to the cover slipping station in the second area.
 3. The apparatus of claim 2, wherein the hoisting device comprises a gripping tool and a lifting tool, the gripping tool configured to grasp the processed microscope slides from the dedicated area on the first area and the lifting tool configured to transfer the processed microscope slides to the cover slipping station on the second area.
 4. The apparatus of claim 1, wherein the conveyor to transport the microscope slides between the plurality of tissue processing stations is configured to carry out movements in a horizontal x-direction and a vertical z-direction.
 5. The apparatus of claim 1, wherein the second area is situated above the first area.
 6. The apparatus of claim 1, wherein the third area is situated below the first area.
 7. The apparatus of claim 1, wherein a pump is connected to each of the reagent supply lines to pump reagent from the reagent storage tank to one or more of the plurality of tissue processing stations.
 8. The apparatus of claim 7, further comprising a filter on each of the reagent supply lines to filter the reagent pumped from the reagent storage tank to one or more of the plurality of tissue processing stations. 